Method to treat diabetes utilizing a gastrointestinal microbiome modulating composition

ABSTRACT

Compositions and methods for treating diabetes are presented herein. In particular, the present disclosure teaches compositions and methods for treating diabetes utilizing a gastrointestinal microbiome modulating composition.

CROSS REFERENCE TO RELATED APPLICATIONS

The present Application is a Continuation Application, which claimspriority to U.S. application Ser. No. 14/238,960, filed on Feb. 14,2014, winch is a U.S. National Stage Application pursuant to 35 U.S.C.§371 of International Application No. PCT/US2012/051408, filed on Aug.17, 2012, which claims priority to U.S. Provisional Application No.61/524,524, filed on Aug. 17, 2011, each of which are herebyincorporated by reference in their entireties.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to weigh control and metabolic fitness.More particularly, the present invention relates to compositions andmethods for increasing desirable microbiota in the human gut andreducing therein undesirable microbiota to achieve a healthy glucoseregulation, metabolic fitness, inclusive intestinal health, and ahealthy body weight.

2. General Background of the Invention

During the past 5 years a large international effort, called the HumanMicrobiome Project (HMP) by the National Institutes of Health, and knownmore broadly as the International Human Microbiome Consortium (IHMC), isaimed at characterizing the microbes living in and on our bodies (seehttp://hmpdace.org/data_browser.php). In the large intestine anestimated 100 trillion microorganisms reside that appear to playessential roles in metabolizing food, drugs, and dietary supplementsthat are not absorbed by the upper gastrointestinal (GI) tract. Inaddition, some of those microorganisms manufacture essential nutrientsand vitamins necessary to sustain health. Such microbial interactions inthe intestinal environment exert critical roles in signaling metabolic-,behavior-, and immune-regulatory systems of the human host.

The first GI microbiomes to be partially characterized are thoseobtained in the feces of obese and lean individuals (1,2). Microbiota ofthe Firmicutes phylum are in abundance in the feces of obese individualswhen compared to that of lean volunteers. Microbiota of the other majordivision, Bacteroidetes, are more abundant than those of Firmicutesdivision in lean individuals. Thus, during metabolic diseases such asobesity and diabetes, an imbalance between these 2 dominant groups ofbacteria reflecting a wealth of Firmuetes species to Bacteroidetesspecies (B:F ratio) is one characteristic of individuals with anunhealthy body mass. These findings are confirmed in obese and lean mice(3). Further, sterile mice can be inoculated with microbiomes obtainedfrom human feces of either obese or lean individuals. When fed astandard mouse chow, they either become obese or remain lean dependingon the source of their GI microbiota transplant (4). Mice transplantedwith human GI microbiota will become obese when fed a chow rich insimple sugars and fat after several week. The B:F ratio of those miceincreased when the diet is switched to one low in both fat and sugar butrich in plain polysaccharides (5,6).

Microbiota belonging to the Bacteriodetes phylum are specialists attransporting and metabolizing carbohydrates (7). They appear to forageon any available carbohydrate in their environment. Most sugars andstarches, however, are metabolized by the host and are absorbed by theupper GI. Indigestible (by the host) carbohydrates do make it to thelower GI where they are consumed by members of the Bacteriodetes. Theend products of this process are small chain fatty acids (SCFAs) thathave health benefits for the host. Many of these indigestiblecarbohydrates are plant polysaccharides or more commonly called fiber orresistant starch and are classified as prebiotics (8). Microbiomescharacterized in the feces of children maintained on a high fiber diethave a greater B:F ratio than those analyzed from feces of childrenconsuming a typical European diet (9).

The follow U.S. and other patent documents are incorporated herein byreference:

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US20090170141 GHRELIN O-ACYLTRANSFERASE (GOAT) BIOCHEMICAL ASSAY

US20100048595 USE OF ARCHAEA TO MODULATE THE NUTRIENT HARVESTINGFUNCTIONS OF THE GASTROINTESTINAL MICROBIOTA

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US20100254949 PREBIOTIC OLIGOSACCHARIDES

US20100317573 PEDIATRIC FIBER MIXTURE

US20110009359 USE OF NON-DIGESTIBLE CARBOHYDRATES FOR IMPROVINGINTESTINAL MICROBIOTA

US20110034407 USE OF SPHINGOMYELIN AND NON-DIGESTIBLE CARBOHYDRATES FORIMPROVING INTESTINAL MICROBIOTA

US20110123504 GUT FLORA AND WEIGHT MANAGEMENT

US20050239706 MODULATION OF FIAF AND THE GASTROINTESTINAL MICROBIOTA ASA MEANS TO CONTROL ENERGY STORAGE IN A SUBJECT

US20110280840 COMPOSITIONS AND METHODS FOR TREATING OBESITY AND RELATEDDISORDERS BY CHARACTERIZING AND RESTORING MAMMALIAN BACTERIAL MICROBIOTA

US20120027724 METHODS OF TREATING IMPAIRED GLUCOSE METABOLISM VIAADMINISTRATION OF ALGAL BIOMASS

US20120058094 COMPOSITIONS AND METHODS FOR TREATING OBESITY AND RELATEDDISORDERS BY CHARACTERIZING AND RESTORING MAMMALIAN BACTERIAL MICROBIOTA

US20120135957 COMPOSITIONS CONTAINING MIXTURES OF FERMENTABLE FIBERS

WO/2012/024638A2 COMPOSITIONS AND METHODS FOR TREATING OBESITY ANDRELATED DISORDERS BY CHARACTERIZING AND RESTORING MAMMALIAN BACTERIALMICROBIOTA

WO/2011/107482A9 METHOD OF DIAGNOSTIC OF OBESITY

WO/2011/103123A2 REDUCING SHORT-CHAIN FATTY ACIDS AND ENERGY UPTAKE INOBESE HUMANS BY MANAGING THEIR INTESTINAL MICROBIAL COMMUNITIES

WO/2011/022660A1 METHODS OF DIAGNOSING AND TREATINGMICROBIOME-ASSOCIATED DISEASE USING INTERACTION NETWORK PARAMETERS

WO/2008/076696A2 THE GUT MICROBIOME AS A BIOMARKER AND THERAPEUTICTARGET FOR TREATING OBESITY OR AN OBESITY RELATED DISORDER

WO/2008/076006A3 THE GUT MICROBIOME AS A BIOMARKER AND THERAPEUTICTARGET FOR TREATING OBESITY OR AN OBESITY RELATED DISORDER

WO/2010/002890A2 METHODS OF PROMOTING WEIGHT LOSS AND ASSOCIATED ARRAYS

WO/2010/002890A3 METHODS OF PROMOTING WEIGHT LOSS AND ASSOCIATED ARRAYS

WO/2010/15842A2 METHODS AND SYSTEMS FOR PHYLOGENETIC ANALYSIS

EP2102350A2 THE GUT MICROBIOME AS A BIOMARKER AND THERAPEUTIC TARGET FORTREATING OBESITY OR AN OBESITY RELATED DISORDER

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BRIEF SUMMARY OF THE INVENTION

The present invention includes a composition and use of a formulation toincrease the ratio of gastrointestinal microbiota in phylumBacteroidetes to microbiota of Firmicutes phylum.

BRIEF DESCRIPTION OF THE DRAWINGS

For a further understanding of the nature, objects, and advantages ofthe prevent invention, reference should be had to the following detaileddescription, read in conjunction with the following drawing, wherein:

FIG. 1 shows fasting blood glucose levels for 60 days of treatment; and

FIG. 2 shows fasting blood glucose levels for 60 days of treatmentdivided into 10 day periods.

DETAILED DESCRIPTION OF THE INVENTION

One aspect of the present invention is to include an indigestiblecarbohydrate such as inulin, oligofructoses, fructo-oligosaccharides,lactulose, galcto-oligosaccharides, xylo-oligosaccharides, resistantstarches, in the formula. In addition to offering food for species ofBacteriodetes and selecting for their growth, production of short chainfatty acids (SCFAs) in the colon will suppress pathogenic bacteria andviruses from invading and colonizing the GI tract, partially inhibithepatic cholesterol biosynthesis, serve as nutrients for colonocytes,reduce colon pH, increase mineral absorption, and serve as ligands tostimulate release of peptide YY (PYY) and glucagon-like peptide-1(GLP-1) into the blood. PYY and GLP-1 are hormones produced and releasedfrom the colon into the blood. PYY signals satiety. GLP-1 stimulates theglucose sensing ability of the insulin secreting cell, prevents thatcell from apoptosis, stimulates proliferation of those pancreatic betacells and inhibits gastric emptying. GLP-1 also stimulates satiety butthis may be due to its important role in decreasing gastric emptying.

Ghrelin is a hormone secreted by the stomach that stimulates appetite.Ghrelin is unique because it must be acylated with a medium chain fattyacid (MCFA) on the 3rd residue to bind and trigger its receptor(GHS-R1a) (10). Ghrelin comprised with octanoic (C-8)- or decanoic(C-10)-MCFAs are the most common forms of active ghrelin (10). There arealso circulating carboxylesterases that cleave the acyl bond linking thefatty acid to the ghrelin backbone and render ghrelin inactive (11) withrespect to appetite and nutrient consumption but may be beneficial forthe treatment of T2DM (12). The acylating enzyme in the stomach cellthat synthesizes ghrelin was identified about 3 years ago (13,14) and isnow a drug target for several groups. The enzyme termed GOAT (ghrelinO-acyltransferase), has a high affinity for MCFAs but can acetylateghrelin with fatty acids of different lengths. Not only does GOAT preferMCFAs, but ghrelin that is acylated with a MCFA is much more potent thanghrelin forced to be synthesized with either a small or long chain fattyacid (15). Interestingly, levels of circulating medium chain fatty acidsat any time point are relatively low when compared to long chain fattyacids. This is primarily a result of diet. Modern diets and most foodsare rich in long chain fatty acids. MCFAs are abundant in coconut oiland human breast milk. Short chain fatty acids (SCFAs) are also rarelyin the food chain; they can be found in small quantities in butter andcoconut. However, they are produced in large quantities by themicroflora in the colon as a byproduct of polysaccharide consumption bythe microbiota of the Bacteriodetes division. Recent evidence indicatesthat GOAT can use a SCFA to acylate ghrelin, rendering the hormoneinactive at its receptor (16).

Thus, another novel aspect of the present invention is a formuladesigned to significantly enhance production or SCFAs by microbiota toserve as substrates for GOAT, render an inactive ghrelin andconsequently decrease appetite.

While aspects of the present invention are aimed at increasing thepopulation of colonic microbiota that garners carbohydrates, there isthe possibility of those microorganisms redirecting theircarbohydrate-harvesting activities from dietary to host polysaccharidesaccording to nutrient availability (7). The colon mucus layer iscomprises of mucins, which are glycoproteins that have O-linkedoligosaccharides or O-glycans, which account for up to 80% of the massof mucin (17). This mucus layer covers the gastrointestinal epitheliumwhere it provides a protective layer from the rich intestinal microbiomeas well as any pathogens. Such engogenous source of glycans in themicrobiome habitat could offer alternative nutrients. However, whendietary glycans are present, it appears that harvesting energy from thediet is preferred (7).

Recently, evidence that bile salts are bacteriostatic to cecalmicrobiota was presented (24). Moreover, the data indicate thatmicrobiota of the Bacteroidetes phylum are most sensitive to bile salts.Secretion of bile salts into the intestinal tract is stimulated by fatingestion. Thus, ingestion of fat containing food increases bilesecretion, which in turn decreases the numbers of Bacteriodetesmicrobiota. Viscous beta-glucan encapsulates or sequesters bile acids(25) and thus has the ability to preserve members of the Bacteroidetes,even when fat is ingested.

Thus, an important aspect of the present invention includes dietaryglycans. Most are indigestible carbohydrates. Since glycans are also inabundance in yeast walls, one source of dietary glycans is from yeast.Other sources include dietary beta-glucan. Beta-glucan is a naturalpolysaccharide that can be isolated from oat, barley and wheat mostcommonly, but also from baker's yeast, certain fungi, mushrooms, andbacteria. Addition of beta-glucan is an important component of thepresent invention to maintain the mucosa protective barrier as well asto offer nutrients to microbiota that prefer to forage on carbohydratesand would increase the B:F ratio.

Another means to alter an imbalance between microbiota belonging toBacteroidetes and Firmicutes phyla is to suppress the growth of membersof one division. Such bacteriostatic or bactericidal actions for membersof Firmicutes division are reported for a number of phenolic compoundsthat are present in large quantities in berries and other fruits (18).Anthocyanins are the predominating group of phenolic compounds presentin berries and are especially abundant (18). A large proportion ofdietary polyphenolics remain unabsorbed in the gut lumen where theybecome concentrated in the ileum and colon. Up to 85% of blueberryanthocyanins enter the colon (19) and appear to exert more growthsuppressing effects on members of the Firmicutes division than theBacteroidetes division (20, 21). Although blueberry powder, blueberrypomace, or blueberry extract containing such polyphenolics is preferred,the present invention preferably includes any fruit or berry ingredientpreparation containing similar phenolics such as those listed inReference 18. The three preferred classes of ingredients, fermentablepolysaccharides, beta-glucans, and berry extracts, can be prepared inany type of edible product such as powder to mix in liquid, a bar,smoothie, yogurt, shake, etc. The blend may also be prepared in acapsule or compressed into a tablet. Preferably, the formulation is tobe ingested twice per day in between the first and second meals as wellas between the second and third meals.

It is preferred that each dose of the formulation contain at least about4 g of indigestible polysaccharide, at least about 2 g beta-glucan andat least about 4 g of berry powder extract (the equivalent of 2 cups (16ounces) or 0.47 liters whole blueberries containing about 800 mg oftotal phenolics. 100 mg of anthocyanins and 6900 μmoles TE antioxidantactivity). It is preferred that the dose be administered twice per daywithin 1 hour prior to meal 1 or within 1 hour prior to meal 2, as wellas within 1 hour prior to meal 3. These are our best estimates but theformulation may change.

Ranges of ingredients preferably include about 2-120 mg/kg of bodyweight of indigestible polysaccharide, about 2-80 mg/kg of body weightof beta-glucan and about 2-120 mg/kg of body weight of blueberrypowdered extract or any fruit or berry ingredient preparation containingsimilar phenolics such as those listed in Reference 18. Ranges of theseingredients more preferably include about 10-80 mg/kg of body weight ofindigestible polysaccharide, about 5-50 mg/kg of body weight ofbeta-glucan and about 10-80 mg/kg of body weight of blueberry powder orany fruit or berry ingredient preparation containing similar phenolicssuch as those listed in Reference 18. Ranges of these ingredients mostpreferably include about 20-60 mg/kg of body weight of indigestiblepolysaccharide that is a fermentable fiber or resistant starch, about10-30 mg/kg of body weight of beta-glucan and about 20-60-320 mg/kg ofbody weight of blueberry powdered extract or any fruit or berryingredient preparation containing similar phenolics such as those listedin Reference 18. One of these ingredients in a formulation is preferableto none, any two are preferable to only one, and most preferably allthree are present in a formulation of the present invention thatcontains less than 70 usable Calories (Cal or keal) (293 Kilojoules).The calorie or joule content of the formulation can be calculated fromderived constants determined for macronutrients in the formula. However,the majority of ingredients in the formula is not digested and absorbedand thus is not assimilated or utilized for energy in the human body. Ausable calorie or joule is only the calorie or joule that is assimilatedor utilized for energy in the human body.

A preferred indigestible carbohydrate is inulin and can be foundnaturally in banana, asparagus, garlic, onions and wheat flour at lowlevels. To consume a sufficient quantity to achieve levels in the mostpreferred dose is possible but that would contribute significant usablecalories or joules to the daily caloric intake of individuals whoalready have an unhealthy weight (see Table below) because the foodsalso contains usable calories or joules. Inulin content of raw Jerusalemartichoke, chicory root, and agave is high. Ingestion of a purifiedpreparation of inulin contributes significantly less usable calories orjoules to daily caloric intake than does ingestion of a vegetablecontaining inulin (see Table 1). To obtain the desired dose of inulin byeating the more common vegetable would contribute about 40-times thecalories or joules that purified inulin contributes.

TABLE 1 Quantity and Calorie or Kilojoule Content for Preferred Dose ofIndigestible Carbohydrate Ingredient Quantity Calories/KilojoulesPurified Inulin 4 g 0.48/2.01 Banana 6 bananas 630/2,638 Raw Asparagus 1cup (0.237 liters) 27/113 Raw Garlic 15 cloves 60/251 Raw Onion 10tablespoons chopped 40/167 Wheat Flour 1 cup (0.237 liters) 455/1,905Raw Jerusalem Artichoke ¼ cup (0.059 liters) 18/75 Raw Chicory Root ½root 20/84

Beta-glucan can be found naturally in oats, barley, mushrooms andBaker's yeast. To consume a sufficient quantity of these foods toachieve levels in the most preferred dose is possible but that wouldcontribute significant useable calories or joules to the daily caloricintake of individuals who already have an unhealthy weight (see Table 2)because the foods also contains usable calories or joules.

TABLE 2 Quantity and Calorie or Kilojoule Content for Preferred Dose ofBeta-Glucan Ingredient Quantity Calories/Kilojoules PromOat Ingredient 2g 5.26/22.02 OatWell 22% 2 g 4.3/18.0 Old Fashion Rolled Oats ½ cup dry(0.118 liters) 150/628 Barley ½ cup dry (0.118 liters) 350/1,465 ShitakeMushrooms 5 cups pieces (1.18 liters) 400/1,675 Baker's Yeast 2 cakes(34 g) 35.8/149.9

Polyphenolics can be divided into major groups of anthocyanidins andflavonoids. They are found naturally in blueberry, cranberry, raspberry,and strawberry. These berries contain different levels of anthocyanidinsand flavonoids according to Reference 22 and the abbreviated tablebelow. Blueberries contain the broadest spectrum of polyphenoliccompounds. Further, ingestion of the equivalent of 2 cups (16 ounces or0.47 liters) of fresh blueberries provides sufficient bioactivecompounds to improve insulin sensitivity in insulin resistant patientsas demonstrated in Reference 23.

TABLE 3 USDA Database for the Anthocyanin Content of Selected Foods,Release 2.1 (2007). Values are the mean in mg/100 g edible portion wholeraw berry. Cyanidin Delphinidin Malvidin Petunidin Anthrocyanin - RawBlueberry 16.97 47.40 61.35 26.42 Anthrocyanin - Raw Cranberry 41.817.66 0.31 0 Anthrocyanin - Raw Raspberry 35.84 0.29 0.70 0Anthrocyanin - Raw Strawberry 1.96 0.32 0 0.08

TABLE 4 USDA Database for the Flavonoid Content of Selected Foods,Release 2.1 (2007). Values are the mean in mg/100 g edible portion wholeraw berry. Epicatechin Catechin Myricetin Quercetin Flavonoid - RawBlueberry 13.69 37.24 2.66 5.05 Flavonoid - Raw Cranberry 4.37 0.39 6.7815.09 Flavoid - Raw Raspberry 4.07 1.56 0 1.23 Flavonoid - RawStrawberry 0.12 3.32 0 1.14

To consume a sufficient quantity (2 cups (16 ounces) or 0.47 liters) toachieve levels in the most preferred dose of blueberry is possible butthat would contribute significant useable calories or joules to thedaily caloric intake of individuals who already have an unhealthy weight(see Table 5) because the food also contains usable calories or joules,especially sugar. Pomace is the solid remains of blueberry afterpressing for juice and thus removes sugar and water when manufacturingberry juice. Pomace contains indigestible carbohydrate that accounts for85% of calories or joules. Pomace can be extracted with solvents such asalcohol or water to remove other useable calories. Blueberry extracts ofpomace contain less than 10 useable Calories (42 Kilojoules).

TABLE 5 Quantity and (Calorie or Kilojoule Content for Preferred Dose ofBerry Pomace Calories (kcal)/(kilojoule) Calories/Kilojoule Berry BerryFresh Whole Berry Pomace (sugar removed) Blueberry (2 cups)  158/662  53.2/223 (0.47 liters) Cranberry (5.38 cups)  323/1,352  179.7/752.4(1.27 liters) Raspberry (9.41 cups)  60/2,520   398/1,666 (2.23 liters)Strawberry (59.3 cups) 2965/12,414 1209.7/5,064.7 (14.03 liters)

In some cases dosing may be once a day to initiate treatment and toacclimate the GI microbiome for up to 1 week and then switched to twicedaily dosing. In other cases twice daily administration of half theoptimal dose is desired such as in children or the elderly. In somecases optimal twice daily dosing may be changed to a daily dosingregimen to maintain the changes stimulated by twice daily dosing. Inother cases the twice daily dosing may be switched to twice daily dosingof half the most preferred dose such as in children or adults who cannottolerate preferred dosing.

The present invention can be used in combination with drugs. Forexample, broad spectrum antibiotics prescribed for systemic infectionsare not completely absorbed by the upper GI system and make it to themicrobiota environment. This will temporarily alter the GI microbiomeand it may be desirable to ingest the formulation of the presentinvention during antibiotic therapy, in preparation for a course ofantibiotics, or following such intervention. Specific antibiotics maytarget Gram positive or Gram negative bacteria. Treatment with a Grampositive antibiotic before-, during-, or following ingestion of theabove combination of prebiotics of the present invention could improvethe desired outcome. Because the formulation of the present invention isexpected to increase GLP-1 levels, it may be desirable to combine thatintervention with a DPP-IV (dipeptidy peptidase type IV) inhibitor suchas Vildagliptin, Sitagliptin, or others in this class. This will greatlyprolong the half-life of GLP-1 released from the colon and thecombination would be preferred over the DPP-IV inhibitor alone or aninjection of a GLP-1 peptide agonist such as Byetta. The presentinvention can also be combined with other diabetes interventions such asbiguanides. An example includes metformin (Glucophage, Glucophage XR,Riomet, Fortamet, and Glumetza). In fact, combined use of the presentinvention with metformin may not only improve blood glucose regulationbut will counter the diarrhea side effect often associated withmetformin (see Example 5). This is a potentially important clinicalobservation. Metformin is recommended as the initial drug of choice forthe treatment of type-2 diabetes (26). Metformin has been reported tocause a 20% incidence of diarrhea in diabetic patients taking the drugcompared to only 6% of diabetic patients not taking metformin (27). Infact, diarrhea with metformin is a sufficient problem that some diabeticpatients cannot tolerate the drug. Since metformin has a good safetyrecord causes an approximate 2-3 kg weight loss and is a low-costgeneric medication, increasing the tolerance to metformin whileincreasing its efficacy using a safe food supplement could havebeneficial public health consequences. The present invention may also becombined with alpha-glucosidase inhibitors such as Precose (acarbose)and Glyset (miglitol) that are effective in controlling blood glucosebut are poorly tolerated because of associated diarrhea. In addition,some drugs such as atypical antipsychotics and insulin cause weight gainin some patients. The present invention could be combined with suchdrugs to combat the weight gain without intervening with antipsychoticefficacy.

The present invention can be used to treat other diseases and syndromes.Irritable bowel syndrome (IBS) is a disorder characterized by abdominalpain or discomfort, and altered bowel habit (chronic or recurrentdiarrhea, constipation, or both—either mixed or in alternation). IBSwith constipation is sometimes referred to as IBS-C orconstipation-predominant IBS. IBS with diarrhea is sometimes referred toas IBS-D. The key symptom of IBS is abdominal pain or discomfortanywhere in the abdomen. It may change over time. The impact of IBS canrange from mild inconvenience to severe debilitation. Persons withmoderate to severe IBS must struggle with symptoms that often impairtheir physical, emotional, economic, educational and social well-being.The exact cause of IBS is not known. Treatments are available for IBS tohelp manage symptoms but none appear to address the etiology. Fecalmicrobiota of patients suffering with IBS have a 2-fold decrease in theratio of the Bacteroidetes to Firmicutes when compared with healthyindividuals (28). Thus, the present invention that stimulates anincrease in the ratio of members of the Bacteroidetes phylum to theFirmicutes phylum should be an efficacious and safe therapeutic to treatIBS.

The present invention can be used in preparation for fecal transplant orfecal bacteriotherapy and its follow up. Fecal microbiotatransplantation is the process of transplantation of fecal microbiotafrom a healthy individual into a recipient as a treatment for diseasessuch as infection by Clostridium difficile (29) or more recently, type 2diabetes (30). Guidelines for both the donor and the recipient are beingprepared and will likely indicate strategies for both the donor and therecipient. The use of the present invention in preparation of bothsubjects for the procedure and continued use by the recipient aftertransplantation will increase the probability of a successful outcome.The present invention can also be used in preparation for fecal banking.Such practice will permit a subject to prepare a specific fecal samplefor banking until needed. This may occur in preparation of prolongedantibiotic surgery, cancer chemotherapy and an unforeseen chronicillness.

The present invention can be used to treat ocular diseases. Recently, itwas demonstrated that the microbiota of a conventional raised laboratorymouse adversely alters the ocular lens (31). The present invention isdesigned to cause a shift in the GI microbiome. Thus, the invention willbe useful in prevention of cataracts. Another eye syndrome is commonlycalled dry-eye syndrome (DES). Traditionally, DES has been thought of asa deficiency of tears at the ocular surface. In recent years, however,investigators have shown that DES is much more complex than previouslythought, and that “tear film dysfunction syndrome” might more accuratelydescribe the condition. Tear film dysfunction can be broken down intotwo basic etiologic classifications: insufficient tear production orincreased evaporation of tears from the eye surface (32). The tear filmis made up of lipid, aqueous and mucin components. Individuals with dryeye syndrome can be deficient in any of these components and all appearto be related, in part, to healthy GI function. Thus, the presentinvention will be useful in treating DES.

TABLE 6 Most Preferred Formulation with Function Ingredient Mass(g/dose) Function Reference Blueberry 4.7 Weight loss, antimicrobial, 1-5 extract antioxidant β-Glucan 2 ↑satiey, ↓LDL chol,↑PYY,  6-11↑GLP-1, ↓ghrelin, ↓hunger Inulin 4 ↑GLP-1, ↑PYY, ↑satiety, ↓ 12-15ghrelin, ↓TG, Support GI Immune System ↑indicates an increase inreported ↓indicates that a decrease is reported TG is triglyceride  1.J. Nutr. 140: 1764-1768, 2010.  2. Chemico-Biological Interactions 189:1-8, 2011  3. J. Agric. Food Chem. 58: 3970, 2010  4. J Appl Microbiol.90: 494, 2001  5. Am J Clin Nutr. 79:727, 2004  6. J Med Food 10: 720,2007  7. Nutr Res 26: 644, 2006.  8. J. Nutr. 138: 732, 2008  9. J Am.Coll Nutr. 26: 639, 2007. 10. Appetite 53: 338, 2009 11. Eur J Clin Nutr62: 600, 2008. 12. Am J Clin Nutr 89:1751, 2009 13. Appl. Physiol. Nutr.Metab. 35: 9-16 (2010) 14. J. Nutr. 137: 2552S-2556S, 2007 15. J. Nutr.137: 2547S-2551S, 2007

TABLE 7 Quantity of Ingredients to Formulate Efficacious DosesContaining No More than 70 Useable Calories (293 Kilojoules) (amount ofinulin and beta glucan are pure - quantities of product containing themwill be higher): Row In- Inulin Beta Glucan Blueberry Pomace 1 gredientfrom Oat Extract Powder Row Al- oligofructoses, Beta-glucan dietary 2ternative fructo-oligosaccharides, from: polyphenolic lactulose, baker'syeast, compounds galcto-oligosaccharides, oat, barley, such as thosearabinoxylans, resistant wheat, fungi present in starch, mushrooms,extracts of xylo-oligosaccharides, bacteria, and pomace polydextrose,soybean other biocompatible from oligosaccharides, isomalto- sourcesberries, oligosaccharides, gluco- grapes, oligosaccharides, palatinose,melons gentio-oligosaccharides, lactitol, sorbitol, maltitol. xylitolRow Function Stimulates production of Decreased total Inhibit growth 3Short chain fatty acids in cholesterol, decrease of some GI colon,suppress pathogenic LDL cholesterol, microbiota induce bacteria andviruses from increase HDL a reduced intestinal colonizing the GI tract,cholesterol, decrease microbiome or serve inhibit hepatic cholesterolblood glucose, protect as an antioxidant, biosynthesis, provideintestinal mucosa, stimulate growth of colonocytes nutrients, prime theGI immune microbiota that thrive reduce colon pH, increase systems,increase in a reduced environment. colon mineral absorption, viscosityof GI provide ligands to stimulate microbiome, absorb release of peptideYY (PYY) excess water in colon. and glucagon-like peptide-1 (GLP-1) intothe blood. produce inactive ghrelin. Row Preferred 1-12 g 0.5-6 g 1-15 g4 Range Row More 2-8 g 0.75-4 g 2-10 g 5 Preferred Range Row Most 3-5 g1-3 g 2-6 g 6 Preferred Range Row Example 4 g 2 g 4.7 g 7Formula A

FORMULA A is made of 4.7 g dried blueberry pomace extract powder fromMilne Fruit Products (Prosser, Wash., product no. FG20155), 9.22 gOatWell 22®beta glucan from CreaNutrition (Sweden), 4.4 g agave inulin(Inufib) from The Iidea Company (Tlaquepaque, Jal., Mexico), withinactive ingredients preferably added for flavor, mouth feel, textureand mixing, including 8.5 g erythritol, 0.5 g soy protein, 0.4 g xanthangum, 0.38 g citric acid, 0.14 g stevia, and 0.4 g natural flavor thatare not believed to affect the efficacy of Formula A. FORMULA A is to beadded to a liquid such as 180 ml of water, milk, juice, etc., mixed andconsumed as a suspension. The dried blueberry pomace extract powder usedin connection with the present invention is preferably blueberry extractpowder, product no. FG20155, available from Milne Fruit Products ofProsser, Wash.

Formula A is used in the examples below. One dose of FORMULA A provides120% of the recommended daily dietary allowance (RDA) of antioxidantactivity for children and adults as well as 30% dietary fiber for menand 50% dietary fiber for women, 60% dietary fiber for children aged1-3, 45% dietary fiber for children aged 4-8, 36% dietary fiber for boysof ages 9-13, and 45% of dietary fiber for girls of ages 9-13.

Preferably a dose of Formula A is taken at least once per day, morepreferably at least twice per day, and even more preferably at least 3times per day. Each dose of FORMULA A is analyzed to contain active andinactive ingredients described in the table 8:

TABLE 8 Active and Inactive Ingredients Measured in the Most PreferredDose of FORMULA A. Analyte Mass/dose Active Ingredients Totalanthrocyanins 162.53 mg Total polyphenolics 723.99 mg Antioxidantactivity 6969.4 μmoles TE Beta glucan 2.03 g Inulin 3.79 g Other solublefiber 3.03 g Insoluble fiber 2.30 g Inactive Ingredients Protein 2.87 gFat 0.34 g Calcium 54.81 mg Iron 1.63 mg Sodium 52.99 mg Potassium132.32 mg Erythritol 8.39 g Sucrose 0.17 g Fructose 1.54 g Glucose &Galactose 0.71 gFurther analysis of the phenolic content of FORMULA A is described bythe table 9:

TABLE 9 Finger Print Analysis of Polyphenolics Present in the MostPreferred Dose of Formula A. Catechins¹ Epigallocatechin Gallate 158.2mg Epicatechin Gallate 110.5 mg Epigallocatechin 21.9 mg Catechin 6.7 mgEpicathecin 3.5 mg Quercetin 3.2 mg Gallocatechin Gallate 1.1 mgGallocatechin ND Catechin Gallate ND Anthocyanins² Malvidin3-O-Galactoside 30.9 mg Malvidin 3-O-Arabinoside 17.7 mg Malvidin3-O-Glucoside 12.4 mg Delphinidin 3-O-Arabinoside 9.4 mg Delphinidin3-O-Galactoside 7.5 mg Petunidin 3-O-Galactoside 6.8 mg Petunidin3-O-Arabinoside 6.4 mg Petunidin 3-O-Glucoside 5.0 mg Delphinidin3-O-Glucoside 4.5 mg Peonidin Chloride 1.5 mg Cyanidin 3-O-Arabinoside1.2 mg Peonidin 3-O-Glucoside 1.2 mg Cyanidin 3-O-Glucoside 0.7 mgDelphinidin Chloride 0.6 mg Peonidin 3-O-Arabinoside 0.5 mg Peonidin3-O-Galactoside 0.4 mg Petunidin Chloride 0.4 mg Cyanidin Chloride 0.4mg Cyanidin 3-O-Galactoside ND Cyanidin 3-O-Rutinoside ND MalvidinChloride ND ¹HPLC anaylsis, ²ORAC trolox equivalents, ND = notdetectable

EXAMPLE 1 Human Study Utilizing FORMULA A to Increase the Ratio ofGastrointestinal Microbiota in Phylum Bacteroidetes to Microbiota ofFirmicutes Phylum, Improve Glucose Regulation and Improve BodyComposition

Subject and Methods

-   a) The required number of subjects are properly screened to fulfill    the necessary qualifications.-   b) appropriate laboratory evaluations are performed,-   c) measures of positive primary and secondary outcome responses are    recorded,-   d) adverse events are documented, and-   e) patients are adequately followed-up.    Overview

This study is expected to demonstrate that overweight subjects withimpaired fasting blood glucose on an ad libitum diet who take FORMULA Aeither within 1 hour prior to meal 1 or within 1 hour prior to meal 2,as well as within 1 hour prior to meal 3 for 4 weeks:

-   -   1. Eliminate stool containing a greater ratio of microbiota        species from the Bacteroidetes phylum to Firmicutes phylum than        this ratio of microbiota in their stool at the start of the        intervention, and when compared to subjects consuming a placebo,        and    -   2. Have an improved oral glucose tolerance test (OGTT) as        measured by blood glucose and insulin levels before, during, and        at 120 minutes after ingestion of 75 g glucose when compared to        their initial OGTT, and when compared to subjects consuming        placebo, and    -   3. Have lower overnight fasting blood glucose levels as measured        by a blood glucose monitor before ingesting a morning meal when        compared to their overnight fasting blood glucose values at the        start of the intervention, and when compared to subjects        consuming placebo,    -   4. Experience an improved body composition as measured by a        decrease in body weight, a decrease in body fat or % body fat, a        decrease in waist circumference measurements, and    -   5. Experience decreased appetite before a meal, increased        satiety during the meal, when compared to subjects consuming        placebo, and    -   6. are found to have elevated GLP-1 as well as PYY levels with        reduced active ghrelin levels after a standardized meal when the        values are compared to those of subjects consuming placebo on        week 3 of the intervention.        General

In this study, subjects consume either 180 ml of FORMULA A formula or aplacebo containing the same total dietary fiber level as FORMULA A butas inactive cellulose orally within 1 hour prior to consumption ofeither meal 1 or meal 2 and within 1 hour prior to consumption of meal 3each day. Placebo formula contains cellulose with food coloring andflavor to match the total dietary fiber content (8.75 g) of FORMULA A.Placebo is prepared by Merlin Development a the same time they prepareFORMULA A

Subjects report weekly for measurements and assessment of any sideeffects. They are asked to collect a stool sample before initiatingeither FORMULA A or placebo intervention as well as at the end of the 4week treatment period. They are also asked to record any side effectsand their frequency (checklist assessment). They are asked to recordappetite (how hungry are you) and satiety (how full are you) during thestandardized meal at the 3^(rd) week of intervention. They are providedwith the proper paper work to record these.

Subject Screening and Selection

A total of 30 subjects is selected, 15 assigned to FORMULA A and 15assigned to placebo.

Qualifications of Subjects

1) Healthy men and women between the ages of 18 and 70 with a BMIbetween 25 and 35 are eligible. 2) Fasting blood glucose between 100 and125 mg/dl. 3) Stable weight over 2 months

Subjects Excluded from the Study

People who:

-   a) take medications affecting glucose,-   b) take medications affecting insulin,-   c) take medications affecting body weight,-   d) take medications affecting bacterial flora,-   e) have intestinal disease or a recent history of intestinal    disease,-   f) have had surgery on stomach or intestine,-   g) are hypothyroid,-   h) are pregnant,-   i) have heart disease.    Appropriate Laboratory Evaluation

Different tests are performed at the screening of potentialparticipants, at the beginning of the study, and at the end of the 4week treatment period.

1) Screening:

Subjects are screened to exclude hypothyroidism, pregnancy, and heartdisease. The following tests can suffice for this: T4 (thyroxin), T3(triiodotyronine), TSH (thyroid stimulating hormone), urine pregnancytest, blood pressure & ECG (electrocardiogram).

2) Beginning of Study:

Subjects passing the initial screen are evaluated at the beginning ofWEEK #1 as follows:

-   -   a) Fasting blood glucose and insulin levels.    -   b) SMA 20 (Sequential multi-channel analysis with computer-20, a        metabolic panel with 20 different analytes), including, uric        acid, and liver function tests    -   c) Triglycerides    -   d) Cholesterol, including fractions    -   e) Glycosylated hemoglobin A1 (HgbA1)    -   f) Weight, taken on the same scale each time    -   g) Body fat % and total body fat, determined by DXA (dual-energy        X-ray absorptiometry).    -   h) Height    -   i) Waist and hip measurements    -   j) Blood glucose, insulin, GLP-1, PYY and ghrelin responses to a        75 g oral glucose challenge    -   k) Assessment of appetite and satiety using a visual analog        scale    -   l) Stool is collected and stored frozen but not analyzed until        the end of study.

3) End of Study Assessment:

-   -   a) all labs and assessments done in step 2 at beginning of        study,    -   b) Analysis of the fecal microbiome DNA from both the initial        sample and the final sample.        Study Design

Subjects selected for participation are allowed an ad libitum diet andare given an evaluation sheet to assess their appetite and satietybefore and after a meal. Foods excluded include alcohol. Low calorie orjoule liquids are stressed in place of high calorie or joule liquidssuch as fruit juices, milk, sweet tea (tea with sugar), regular softdrinks, coffee with sugar, etc. The subjects are randomly assigned toeither FORMULA A or placebo treatments. Both the experimenter and thesubjects are blinded to who receive FORMULA A or the placebo. Thesubjects are encouraged to consume either treatment within 1 hour priorto either breakfast or lurch and within 1 hour prior to dinner.

Duration

Subject are given a 4 weeks supply of either FORMULA A or placebo at theonset and are instructed to consume the entire 180 ml volume within 1hour prior to either meals 1 or 2, as well as another 180 ml volumecontaining either FORMULA A or placebo within 1 hour prior to meal 3. Adlibitum diets are followed for 4 weeks, but the volunteers areinstructed to consume either FORMULA A or placebo as their only betweenmeal snack.

Outcome

This study is expected to demonstrate that FORMULA A:

-   -   1) Increases the ratio Bacteroidetes species to Firmicutes        species in fecal samples when samples at the end of study are        compared to samples at the onset of study; and when the ratio of        species is compared to samples from the placebo group    -   2) Improves the blood glucose and insulin responses to an OGTT        by decreasing the areas under the insulin curve (improved        insulin sensitivity);    -   3) Decrease fasting blood glucose values    -   4) Produces weight loss, loss of body fat, and (or) decrease of        body fat %    -   5) Increases GLP-1 and PYY response to the oral glucose        challenge and decreases the fasting ghrelin levels at 1 hour        after the both the OGTT and the standardized meal when comparing        final values to the initial measurements of the OGTT, and when        comparing to those findings of the placebo group during the        standardized meal; and    -   6) Decreases stool pH;    -   7) Increases stool SCFA.

If subjects took FORMULA A for periods longer than 4 weeks, particularlyfor at least 8 weeks, the subjects would experience significant weightloss that was primarily fat loss.

EXAMPLE 2 Human Study Utilizing Either FORMULA A or a Placebo SnackReplacement to Increase the Ratio of Gastrointestinal Microbiota inPhylum Bacteroidetes to Microbiota of Firmicutes Phylum, Improve GlucoseRegulation and Improve Body Composition

Subject and Methods

-   a) The required number of subjects are properly screened to fulfill    the necessary qualifications,-   b) appropriate laboratory evaluations are performed,-   c) measures of positive primary and secondary outcome responses are    recorded,-   d) adverse events are documented, and-   e) patients are adequately followed-up.    Overview

This study is expected to demonstrate that overweight subjects withimpaired fasting blood glucose on an ad libitum diet who take FORMULA A,but not those assigned to consume a placebo, within 2 hours prior toconsuming either meal 1 or meal 2, as well as within 2 hours prior toconsuming meal 3 as a snack replacement for 4 weeks:

-   -   1. Eliminate stool containing a greater ratio of microbiota        species from the Bacteroidetes phylum to Firmicutes phylum than        this ratio of microbiota in their stool at the start of the        intervention, and    -   2. Have an improved oral glucose tolerance test (OGTT) as        measured by blood glucose and insulin levels before, during, and        at 120 minutes after ingestion of 75 g glucose when compared to        their initial OGTT, and    -   3. Experience an improved body composition as measured by a        decrease in body weight, a decrease in body fat or % body fat, a        decrease in waist circumference measurements, and    -   4. Experience lower fasting blood glucose levels when compared        to either the initial values or those of the placebo group, and    -   5. Experience decreased appetite before a meal, increased        satiety during the meal, and    -   6. are found to have elevated GLP-1 as well as PYY levels with        reduced active ghrelin levels at 1 hour after both the OGTT and        the standardized meal when the values are compared to those at        the initiation of the trial (OGTT) or the placebo group        (standardized meal test).        General

In this study, subjects are randomly selected to consume 180 ml ofeither FORMULA A formula or placebo formula orally within 2 hours priorto ingestion of either meals 1 or 2 as well as within 2 hours prior toconsumption of meal 3 each day. The subjects and experimenters areblinded to the treatment assignments. Placebo formula contains cellulosewith food coloring and flavor to match the total dietary fiber content(8.75 g) of FORMULA A. Placebo is prepared by Merlin Development at thesame time they prepare FORMULA A. Both formulations are coded by MerlinDevelopment and the code is maintained with them as well as is held inconfidence by a pharmacist at the study clinic until all data arecollected at the end of study.

Subjects report weekly for measurements and assessment of any sideeffects. They are asked to collect a stool sample before initialingFORMULA A or Placebo intervention as well as at the end of the 4 weektreatment period. They are also asked to record any side effects andtheir frequency (checklist assessment). They are asked to recordappetite (how hungry are you) and satiety (how full are you) during theOGTT at both the onset and at the end of the trial as well as during a3^(rd) standardized meal. They are provided with the proper paper workto record these.

Subject Screening and Selection

A total of 30 subjects is selected. Fifteen subjects are randomlyassigned to consume FORMULA A and the remaining 15 subjects are assignedto the FORMULA A group. The demographics of each group are carefullymatched.

Qualifications of Subjects

1) Healthy men and women between the ages of 18 and 70 with a BMIbetween 25 and 35 are eligible. 2) Fasting blood glucose between 100 and125 mg/dl. 3) Stable weight over 2 months

Subjects Excluded from the Study

People who:

-   a) take medications affecting glucose,-   b) take medications affecting insulin,-   c) take medications affecting body weight,-   d) take medication affecting bacterial flora,-   e) have intestinal disease or a recent history of intestinal    disease,-   f) have had surgery on stomach or intestine,-   g) are hypothyroid,-   h) are pregnant,-   i) have heart disease.    Appropriate Laboratory Evaluation

Different tests are performed at the screening of potentialparticipants, at the beginning of the study, and at the end of the 4week treatment period.

4) Screening:

Subjects are screened to exclude hypothyroidism, pregnancy, and heartdisease. The following tests can suffice for this: T4 (thyroxin), T3(triiodotyronine), TSH (thyroid stimulating hormone), urine pregnancytest, blood pressure & ECG (electrocardiogram).

5) Beginning of Study:

Subjects passing the initial screen are evaluated at the beginning ofWEEK #1 as follows:

-   -   a) Fasting blood glucose and insulin levels.    -   b) SMA 20 (Sequential multi-channel analysis with computer-20, a        metabolic panel with 20 different analytes), including, uric        acid, and liver function tests    -   c) Triglycerides    -   d) Cholesterol, including fractions    -   e) Glycosylated hemoglobin A1 (HgbA1)    -   f) Weight, taken on the same scale each time    -   g) Body fat % and total body fat, determined by DXA (dual-energy        X-ray absorptiometry).    -   h) Height    -   i) Waist and hip measurements    -   j) Blood glucose, insulin, GLP-1, PYY and ghrelin responses to a        75 g oral glucose challenge    -   k) Assessment of appetite and satiety using a visual analog        scale    -   l) Stool is collected and stored frozen but not analyzed until        the end of study.

6) End of Study Assessment:

-   -   c) all labs and assessments done in step 2 at beginning of        study,    -   d) Analysis of the fecal microbiome DNA from both the initial        sample and the final sample.        Study Design

Subjects selected for participation are allowed an ad libitum diet andare given an evaluation sheet to assess their appetite and satietybefore and after a meal. Foods excluded include alcohol. Low calorie orjoule liquids are stressed in place of high calorie or joule liquidssuch as fruit juices, milk, sweet tea (tea with sugar), regular softdrinks, coffee with sugar, etc. The subjects are encouraged to consumeeither FORMULA A or Placebo as their only between meal snack.

Duration

Subjects are given a 4 weeks supply of ether FORMULA A or Placebo at theonset and are instructed to consume the entire 180 ml volume containingeither formula within 2 hours prior to either meals 1 or 2, as well asanother 180 ml volume containing either formula within 2 hours prior tomeal 3. Ad libitum diets are followed for 4 weeks, but the volunteersare instructed to consume either FORMULA A or Placebo as their onlybetween meal snack.

Outcome

This study is expected to demonstrate that FORMULA A and not Placebo:

-   -   1) Increases the ratio of Bacteroidetes species to Firmicutes        species in fecal samples when samples at the end of study are        compared to samples at the onset of study;    -   2) Improves the blood glucose and insulin responses to an OGTT        by decreasing the areas under the insulin curve (improved        insulin sensitivity);    -   3) produces weight loss, loss of body fat, and (or) decrease of        body fat %;    -   4) Increases GLP-1 and PYY response to the oral glucose        challenge and decreases the fasting ghrelin levels prior to the        OGTT when comparing final values to the initial measurements;    -   5) Decreases stool pH;    -   6) Increases stool SCFA.

If subjects took FORMULA A as a snack replacement for periods longerthan 4 weeks, particularly for at least 8 weeks, the subjects wouldexperience significant weight loss that was primarily fat loss.

EXAMPLE 3 Human Study Utilizing FORMULA A in Combination with anInhibitor of Dipeptidyl Peptidase-4 (DPP-4) to Increase the Ratio ofGastrointestinal Microbiota in Phylum Bacteroidetes to Microbiota ofFirmicutes Phylum and Improve Glucose Regulation by Sustained Elevationof GLP-1

Subject and Methods

-   a) The required number of subjects are properly screened to fulfill    the necessary qualifications,-   b) appropriate laboratory evaluations are performed,-   c) measures of positive primary and secondary outcome response are    recorded,-   d) adverse events are documented, and-   e) patients are adequately followed-up.    Overview

This study is expected to demonstrate that Type 2 diabetic (T2D)subjects with insulin resistance on an ad libitum diet who take a DPP-4inhibitor and FORMULA A within 1 hour prior to either meal 1 or meal 2,as well as within 1 hour prior to meal 3 for 4 weeks:

-   -   1. Eliminate stool containing a greater ratio of microbiota        species from the Bacteroidetes phylum to Firmicutes phylum than        this ratio of microbiota in their stool at the start of the        intervention and when compared to those only taking the DPP-4        inhibitor, and    -   2. Have improved insulin sensitivity when compared to both        initiation of the study and when compared to those only taking a        DPP-4 inhibitor. Insulin sensitivity is measured by an oral        glucose tolerance test (OGTT). This is performed by measuring        blood glucose and insulin levels before, during, and at 120        minutes after ingestion of 75 g glucose when compared to their        initial OGTT, and    -   3. Have improved fasting blood glucose values when compared to        those only taking the DPP-4 inhibitor, and    -   4. experience an improved body composition as measured by a        decrease in body weight, a decrease in body fat or % body fat, a        decrease in waist circumference measurements when compared to        their baseline values and when compared to those only taking the        DPP-4 inhibitor, and    -   5. experience decreased appetite before a standardized meal,        increased satiety during that meal when compared to their        baseline values and when compared to those only taking the DPP-4        inhibitor, and    -   6. are found to have elevated GLP-1 as well as PYY levels with        reduced ghrelin levels at 1 hour after the both the OGTT and the        standardized meal when the values are compared to their baseline        values and when compared to those only taking the DPP-4        inhibitor.        General

In this study, T2D patients are randomly assigned to either consume 180ml of FORMULA A or a placebo formula containing cellulose orally within1 hour prior to either meals 1 or 2 as well as within 1 hour prior tomeal 3 each day. Patients and experimenters are blinded to thisassignment. All patients are also instructed to take sitagliptin(Januvia®) at the recommended dose of 100 mg, once per day in themorning prior to meal 1 as a treatment to manage their diabetes.

Subjects report weekly for measurements and assessment of any sideeffects. They are asked to collect a stool sample before the initiationof the trial as well as at the end of the 4 week treatment period. Theyare also asked to record any side effects and their frequency (checklistassessment). They are asked to record appetite (how hungry are you) andsatiety (how full are you) during the OGTT at both the onset and at theend of the trial as well as before and during a standardized meal at the3^(rd) week of treatments. They are provided with the proper paper workto record these.

Subject Screening and Selection

A total of 24 subjects is selected. 12 will be randomly assigned toreceive Januvia®+placebo (a cellulose solution that contains the sametotal dietary fiber content as FORMULA A and mimics FORMULA A in colorand taste) or Januvia®+FORMULA A.

Qualifications of Subjects

1) T2D men and women between the ages of 18 and 70 with a BMI between 25and 35 are eligible. 2) Fasting blood glucose between greater than 125mg/dl. 3) Stable weight over 2 months

Subjects Excluded from the Study

People who:

-   a) take medications affecting glucose other than Januvia,-   b) take medications affecting insulin other than Januvia,-   c) take medications affecting body weight,-   d) take medications affecting bacterial flora,-   e) have intestinal disease or a recent history of intestinal    disease,-   f) have had surgery on stomach or intestine,-   g) are hypothyroid,-   h) are pregnant,-   i) have heart disease.    Appropriate Laboratory Evaluation

Different tests are performed at the screening of potentialparticipants, at the beginning of the study, and at the end of the 4week treatment period.

7) Screening:

Subjects are screened to exclude hypothyroidism, pregnancy, and heartdisease. The following tests can suffice for this: T4, T3, TSH, urinepregnancy test, blood pressure & ECG. Fasting blood glucose, fastinginsulin and HgbA1 levels are also measured as an assessment of theirdiabetic state.

8) Beginning of Study:

Subjects passing the initial screen are evaluated at the beginning ofWEEK #1 as follows:

-   -   a) Fasting blood glucose, insulin, and HgbA1 levels.    -   b) SMA 20, including uric acid and liver function tests    -   c) Triglycerides    -   d) Cholesterol, including fractions    -   f) Weight, taken on the same scale each time    -   g) Body fat % and total body fat, determined by DXA.    -   h) Height    -   i) Waist and hip measurements    -   j) Blood glucose, insulin, GLP-1, PYY and ghrelin responses to a        75 g oral glucose challenge    -   k) Assessment of appetite and satiety before, during and after a        standardized meal using a visual analog scale    -   l) Stool is collected and stored frozen but not analyzed until        the end of study.

9) End of Study Assessment:

-   -   e) all labs and assessments done in step 2 at beginning of        study,    -   f) Analysis of the fecal microbiome DNA from both the initial        sample and the final sample.        Study Design

Patients selected for participation are allowed an ad libitum diet andare given an evaluation sheet to assess their appetite and satiety.Foods excluded include alcohol. Low calorie or joule liquids arestressed in place of high calorie or joule liquids such as fruit juices,milk, sweet tea (tea with sugar), regular soft drinks, coffee withsugar, etc. All 24 patients are also instructed to take sitagliptin(Januvia®) at the recommended dose of 100 mg, once per day in themorning with or without food as a treatment to manage their diabetes. 12patients are randomly selected to also consume FORMULA A before 2 of 3daily meals and the remaining 12 patients are instructed to consume aplacebo before 2 of 3 daily meals. Patients and investigators areblinded to whether the snack replacement is placebo or FORMULA A.

Duration

Subjects are given a 4 weeks supply of sitagliptin and either FORMULA Aor placebo at the onset and are instructed to consume the entire 180 mlvolume of either snack replacement within 1 hour prior to either meals 1or 2, as well as another 180 ml volume of snack replacement within 1hour prior to meal 3. All subjects are required to take 1 tablet ofsitagliptin daily (100 mg) in the morning with or without food. Adlibitum diets are followed for 4 weeks.

This study is expected to demonstrate that FORMULA A:

-   -   1) Increases the ratio of Bacteroidetes species to Firmicutes        species in fecal samples when samples at the end of study from        those assigned to FORMULA A are compared to samples at the onset        of study and when subjects taking Januvia+FORMULA A are compared        to patients taking Januvia+placebo.    -   2) Improves the blood glucose and insulin responses to an OGTT        by decreasing the areas under the insulin curve (improved        insulin sensitivity) when subjects taking Januvia+FORMULA A are        compared to patients taking Januvia+placebo    -   3) produces weight loss, loss of body fat, and (or) decrease of        body fat % when patients assigned to FORMULA A are compared to        samples at the onset of study and when subjects taking        Januvia+FORMULA A are compared to patients taking        Januvia+placebo.    -   4) produces decreased fasting blood glucose levels when subjects        taking Januvia+FORMULA A are compared to patients taking        Januvia+placebo, and    -   5) increases GLP-1 and PYY response to the oral glucose        challenge and decreases the fasting ghrelin levels prior to the        OGTT when patients assigned to FORMULA A are compared to samples        at the onset of study and when subjects taking Januvia+FORMULA A        are compared to patients taking Januvia+placebo,    -   6) Decreases stool pH when patients assigned to the FORMULA A        arm are compared to samples at the onset of study and when        subjects taking Januvia+FORMULA A are compared to patients        taking Januvia+placebo    -   7) Increases stool SCFA when patents assigned to the FORMULA A        arm are compared to samples at the onset of study and when        subjects taking Januvia+FORMULA A are compared to patients        taking Januvia+placebo

If subjects took FORMULA A with other DPP-IV inhibitors or otherformulations of Januvia, the subjects are expected to also havesignificantly improved glucose regulation.

EXAMPLE 4 Study Utilizing FORMULA A Snack Replacement to Increase theRatio of Gastrointestinal Microbiota in Phylum Bacteroidetes toMicrobiota of Firmicutes Phylum, Improve Glucose Regulation and ImproveBody Composition in Overweight Children

Subject and Methods

-   a) The required number of children are properly screened to fulfill    the necessary qualifications and their parental consent is obtained,-   b) appropriate laboratory evaluations are performed,-   c) measures of positive primary and secondary outcome responses are    recorded,-   d) adverse events are documented, and-   e) children and their parents are adequately followed-up.    Overview

This study is expected to demonstrate that overweight children withprediabetes or at high risk of developing T2D (type 2 diabetes) on an adlibitum diet who take FORMULA B (identical active ingredients to FORMULAA but formulated in a child friendly delivery system such as ice cream,jelled animals, cookies, etc.) within 1 hour prior to either meal 1 ormeal 2, as well as within 1 hour prior to meal 3 for 4 weeks:

-   -   1. Eliminate stool containing a greater ratio of microbiota        species from the Bacteroidetes phylum to Firmicutes phylum than        this ratio in their stool at the start of the intervention, and    -   2. Have an improved oral glucose tolerance test (OGTT) as        measured by blood glucose and insulin levels before, during, and        at 120 minutes after ingestion of 1.75 g glucose/kg body weight        up to 75 g glucose when compared to their initial OGTT, and    -   3. experience an improved body composition as measured by a        decrease in body weight, a decrease in body fat or % body fat, a        decrease in waist circumference measurements, and    -   4. Experience decreased fasting blood glucose levels    -   5. experience decreased appetite before a meal, increased        satiety during the meal, and    -   6. are found to have elevated GLP-1 as well as PYY levels with        reduced ghrelin levels 1 hour after the OGTT when the values are        compared to those at the initiation of the trial.        General

In this study, children consume 6 jelled animals of FORMULA B formula(each jelled animal contains about 20 g of FORMULA B) within 1 hourprior to either meals 1 or 2 as well as within 1 hour prior to meal 3each day.

Subjects report weekly for measurements and assessment of any sideeffects. They are asked to collect s stool sample before initiatingFORMULA B intervention as well as at the end of the 4 week treatmentperiod. They are also asked to report side effects to their parents whorecord them and their frequency (checklist assessment). The parents areinstructed to ask and to record appetite (how hungry are you) andsatiety (how full are you) before, during, and after a standardized3^(rd) meal at the beginning of study and at the end. The investigatorsscore the same assessment during the OGTT at both the onset and at theend of the trial as well as at home. The parents are provided with theproper paper work to record these.

Subject Screening and Selection

A total of 10 children is selected.

Qualifications of Subjects:

1) Healthy prepubertal boys and girls between the ages of 7 and 12 witha BMI between 25 and 30 are eligible. 2) Fasting blood glucose between100 and 125 mg/dl.

Subjects Excluded from the Study

Children who:

-   a) take medications affecting glucose,-   b) take medications affecting insulin,-   c) take medications affecting body weight,-   d) take medications affecting bacterial flora.-   e) have intestinal disease or a recent history of intestinal    disease,-   f) have had surgery on stomach or intestine,-   g) are hypothyroid,    Appropriate Laboratory Evaluation

Different tests are performed at the screening of potentialparticipants, at the beginning of the study, and at the end of the 4week treatment period.

Screening:

Children are screened to exclude hypothyroidism and puberty. Thefollowing tests can suffice for this: T4, T3, TSH, a physical exam, andin questionable cases based on the physical exam or peripubertalpresentations, a gonadotropin-releasing hormone challenge test.

Beginning of Study:

Children passing the initial screen are evaluated at the beginning ofWEEK #1 as follows:

-   a) Fasting blood glucose and insulin levels.-   b) SMA 20, including uric acid and liver function tests-   c) Triglycerides-   d) Cholesterol, including fractions-   e) Glycosylated hemoglobin A1 (HgbA1)-   f) Weight, taken on the same scale each tine-   g) Body fat % and total body fat, determined by DXA.-   h) Height-   i) Waist and hip measurements-   j) Blood glucose, insulin, GLP-1, PYY and ghrelin responses to a    1.75 g/kg (up to 75 g) oral glucose challenge-   k) Assessment of appetite and satiety using a visual analog scale-   (l) Stool is collected and stored frozen but not analyzed until the    end of study.    End of Study Assessment:    -   a) all labs and assessments done in step 2 at beginning of        study,    -   b) Analysis of the fecal microbiome DNA from both the initial        sample and the final sample.        Study Design

Children selected for participation are allowed an ad libitum diet andtheir parents are given an evaluation sheet to assess their appetite andsatiety. Low calorie or joule liquids are stressed in place of highcalorie or joule liquids such as fruit juices, milk, regular softdrinks, coffee with sugar, etc. The children are encouraged to consumeFORMULA B as their only between meal snack. Other snacks such as candy,ice cream, milk shakes, cookies, potato chips, etc. are discouraged.

Duration

Children are given a 4 weeks supply of FORMULA B at the onset and areinstructed to consume the entire 6 jelled animals containing FORMULA Bwithin 1 hour prior to either meal 1 or 2, as well as another 6 jelledanimals containing FORMULA B within 1 hour prior to meal 3. Ad libitumdiets are followed for 4 weeks, but the children are instructed toconsume FORMULA B as their only between meal snack.

Outcome

This study is expected to demonstrate that FORMULA B:

-   -   1) Increases the ratio of Bacteroidetes species to Firmicutes        species in fecal samples when samples at the end of study are        compared to samples at the onset of study.    -   2) Improves the blood glucose and insulin responses to an OGTT        by decreasing the areas under the insulin curve (improved        insulin sensitivity)    -   3) produces weight loss, loss of body fat, and (or) decrease of        body fat %    -   4) Increases GLP-1 and PYY response to the oral glucose        challenge and decreases the fasting ghrelin levels prior to the        OGTT when comparing final values to the initial measurements.    -   5) Decreases stool pH    -   6) Increases stool SCFA

If children took FORMULA A for periods longer than 4 weeks as a snackreplacement, particularly for at least 8 weeks, the children wouldexperience significant weight loss that was primarily fat loss.

EXAMPLE 5 Formula A Augments the Efficacy and GastrointestinalTolerability of Metformin: A Case report

Subject

JH was a 30 year old Caucasian male who gained 10 kg over the course of8 months and developed lower back pain. He presented to his primaryphysician with that complaint.

The patient was taking omeprazole (20 mg/d), lisinopril (40 mg/d),metoprolol (100 mg/d), and hydrochlorothiazide (25 mg/d) at the time ofhis presentation, had no allergies and had undergone no surgeries. Hedenied past hospitalizations, trauma with residua or blood transfusions.He had never smoked, drank 3-4 alcoholic drinks per week on average andhad no history of intravenous drug use. He was a full-time law studentand had no exposures to occupational toxins. During the 6 months priorto his presentation he had been sexually active with one woman. Thepatient's mother and father were living and well. He had one sister whowas living and well, and he had no children. There was no family historyof diabetes. He had no complaint other than lower back pain. On physicalexamination he was 104 kg with a height of 165 cm and a body mass index(BMI) of 38.3 kg/m². His blood pressure was 116/70 with a pulse rate of80/min and he was afebrile. Urinalysis demonstrated presence of glucose,that was followed up by a non-fasting capillary blood glucose of 450mg/dl glucose and a hemoglobin A1c of 8.8% and a microalbumin/creatineratio of 147. His physician made the diagnosis of type 2 diabetesmellitus based on his age, lack of acidosis and a fasting blood sugar of375 mg/dL. He was started on metformin 500 mg twice a day and wasinstructed to measure his fasting blood sugar on a daily basis. Over thecourse of 7 days he was instructed to increase metformin to 1000 mgtwice a day. Over the course of the next 9 days he developed persistentwatery stools and his fasting blood sugar decreased to about 325 mg/dL.He knew from his contact with NuMe Health, LLC that the company wasdeveloping and testing a dietary supplement with a goal of increaseinsulin sensitivity through a salutary effect on the stool microbiome.On the 9th day of his treatment with 1000 mg metformin dose, he addedFORMULA A taken twice a day within an hour before breakfast or lunch andwithin an hour before dinner. Within two days of starting FORMULA A hisfasting glucose dropped from 325 mg/dL to 175 mg/dL and after 5 weeks oftaking the FORMULA A along with the metformin, his fasting blood glucosewas 100 mg/dL (FIG. 1).

FIG. 1 shows fasting blood glucose levels at the start of 1000 mgmetformin twice daily. Formula A was added twice daily on day 9 and thecombination was continued except from days 49 to 51 when daily dosing ofFORMULA A wax missed. Diarrhea was associated with metformin except whenFormula A was added.

Dividing the first 60 days of diabetes treatment into 10 day periods,his blood sugar and its variance (standard deviation) decreased from344±44.1 mg/dL in the first 10 days, to 182.2±14.7 in the second 10days, 144.3±9.6 during the third 10 days, 114.5±11.8 during the fourth10 days, 123.0±9.6 during the fifth 10 days, and 121.0±11.8 during thesixth 10 days (FIG. 2).

During this period, his weight decreased by 5.5 kg, and his waterystools reverted to soft regular formed bowel movements. After anadditional month, he ran out of his FORMULA A for 2 days. During thatperiod, his blood sugar increased from 100 mg/dL to 131 mg/dL and hisdiarrhea returned. After resuming FORMULA A his blood sugar returned to113 mg/dL and his stools return to being formed and soft. His diabeticcontrol and stools have remained under control on the combination ofmetformin and FORMULA A at the time of this writing and he has lostabout 5.5 kg body weight. His hemoglobin A1c decreased to 6.3% and amicroalbumin/creatine ratio was 16.

Discussion

This case report is encouraging. Not only did FORMULA A appear to have apositive effect on fasting blood sugar, but the watery stools induced bytreatment with metformin were normalized. This is a potentiallyimportant clinical observation. Metformin has been recommended as theinitial drug of choice for the treatment of type-2 diabetes (26).Metformin has been reported to cause a 20% incidence of diarrhea indiabetic patients taking the drug compared to only 6% of diabeticpatients not taking metformin (27). In fact, diarrhea with metformin isa sufficient problem that some diabetic patients cannot tolerate thedrug. Since metformin has a good safety record, causes an approximate2-3 kg weight loss and is a low-cost generic medication, increasing thetolerance to metformin while increasing its efficacy using a safe foodsupplement could have beneficial public health consequences.

Acarbose is another drug used to treat diabetes that is associated withthe side effect of a watery stool because of its mechanisms of action.If FORMULA A is used in conjunction with acarbose to treat type 2diabetes, both better control of blood glucose and solid stool formationare expected.

EXAMPLE 6 Human Study Utilizing Either FORMULA A or a Placebo toIncrease the Ratio of Gastrointestinal Microbiota in PhylumBacteroidetes to Microbiota of Firmicutes Phylum and Improve Symptoms ofIrritable Bowel Syndrome (IBS)

Subject and Methods

-   a) The required number of subjects are properly screened to fulfill    the necessary qualifications,-   b) appropriate laboratory evaluations are performed,-   c) measures of positive primary and secondary outcome responses are    recorded,-   d) adverse events are documented, and-   e) patients are adequately followed-dup.    Overview

This randomized, placebo-controlled clinical trial is expected todemonstrate the efficacy and tolerability of FORMULA A indiarrhea-predominant humans with IBS. Subjects assigned to consumeFORMULA A but not a placebo, within 1 hour prior to consuming eithermeal 1 or meal 2, as well as within 1 hour prior to consuming meal 3 for4 weeks:

-   -   1. Eliminate stool containing a greater ratio of microbiota        species from the Bacteroidetes phylum to Firmicutes phylum, and    -   2. Report adequate relief for all 4 weeks, and    -   3. Report decreased urgency, and    -   4. Report decreased stool frequency, and    -   5. Have firm stools within 1 week of starting treatment.        General

In this study, subjects are randomly selected to consume 180 ml ofeither FORMULA A formula or placebo formula orally within 1 hour priorto ingestion of either meals 1 or 2 as well as within 1 hour prior toconsumption of meal 3 each day. The subjects and experimenters areblinded to the treatment assignments. Placebo formula contains cellulosewith food coloring and flavor to match the total dietary fiber content(8.72 g) of FORMULA A. Placebo is prepared by Merlin Development at thesame time they prepare FORMULA A. Both formulations are coded by MerlinDevelopment and the code is maintained with them as well as is held inconfidence by a pharmacist at the study clinic until all data arecollected at the end of study.

Subjects report weekly for measurements and assessment of any sideeffects. They are asked to collect a stool sample before initiatingFORMULA A or Placebo intervention as well as at the end of the 4 weektreatment period. During the screening, treatment, and follow-upperiods, daily and weekly symptom data are collected using aninteractive telephone-based system.

Pain and bowel function data are collected during the screening phase toensure that patients had a suitable symptom level at study entry.Severity of pain and discomfort was assessed daily on a 5-point scale(0, none; 1, mild; 2, moderate; 3, intense; and 4, severe). Stoolconsistency data are monitored daily and scored as follows: 1, veryhard; 2, hard; 3, formed; 4, loose; and 5, watery. Absence of stool wasassigned a value of 0. Patients also record their IBS symptoms urgency(0%, feel no need to evacuate-100%, feel severe need to evacuate), stoolfrequency (# of stools per day), bloating (0, no sensation of extendedabdomen; 1, mild; 2, moderate; 3, severe) and sense of incompleteevacuation (0, sensation of complete evacuation; 1, incomplete; 2,constipated) daily during the treatment and follow-up phases.

Subject Screening and Selection

Patients with IBS and a diarrhea-predominant bowel pattern aged 18 yearsor older are enrolled in this study if their symptoms fulfilled the RomeI criteria for IBS for at least 6 months. Patients undergo a 2-weekscreening evaluation to confirm sufficient level of pain and stoolconsistency before randomization. Since no objective criteria exist forsubgrouping of IBS patients, physicians are asked to assess patientsaccording to predominant pattern of bowel function based on thepatient's disease history. Physicians are provided with a guidelinebased on the percentage of time the patient had experienced diarrhea. Ifdiarrhea is present for ≧75% of the time, then the patient is classifiedas being diarrhea predominant.

Patients are excluded if they are pregnant, breastfeeding, or not usingapproved methods of contraception (if of child-bearing potential); if anunstable medical or other gastrointestinal condition exists; if there isa major psychiatric disorder or substance abuse within the previous 2years; if an investigational drug was used within 30 days of thescreening phase; or if a prohibited concurrent medication (likely tointerfere with gastrointestinal tract function or analgesia) was usedwithin 7 days before entering the screening phase.

Pain and bowel function data are collected during the screening phase toensure that patients has a suitable symptom level at study entry asdescribed above.

Appropriate Symptom and Laboratory Evaluation

Evaluations are performed at the screening of potential participants, atthe beginning of the study, daily, and at the end of the 4 weektreatment period.

1) Beginning of Study and Daily:

-   -   a) Severity of pain and discomfort is assessed on a 5-point        scale (0, none; 1,. mild; 2, moderate; 3, intense; and 4,        severe).    -   b) Stool consistency data are scored as follows: 1, very hard;        2, hard; 3, formed; 4, loose; and 5, watery. Absence of stool        was assigned a value of 0.    -   c) Urgency (0%, feel no need to evacuate-100%, feel severe need        to evacuate).    -   d) Stool frequency (# of stools per day)    -   e) Bloating (0, no sensation of extended abdomen; 1, mild; 2,        moderate; 3, severe)    -   f) Sense of incomplete evacuation (0, sensation of complete        evacuation; 1, incomplete; 2, constipated)    -   g) Body weight.

2) Beginning of Study:

Stool is collected and stored frozen but not analyzed until the end ofstudy.

3) End of Study Assessment:

Analysis of the fecal microbiome DNA from both the initial sample andthe final sample.

Study Design

Subjects selected for participation are allowed an ad libitum diet.Foods excluded include alcohol. The subjects are encouraged to consumeeither FORMULA A or Placebo within 1 hour prior to 2 meals each day withingestion of the test agent being mandatory prior to the 3^(rd) meal.

Subjects are given a 4 week supply of ether FORMULA A or Placebo at theonset and are instructed to consume the entire 180 ml volume containingeither formula within 1 hour prior to either meals 1 and 2, as well asanother 180 ml volume containing either formula within 1 hour prior tomeal 3.

Subjects report weekly for measurements and assessment of IBS symptoms.During the screening and treatment periods, daily symptom data arecollected using an interactive telephone-based system.

Outcome

This study is expected to demonstrate that FORMULA A and not Placebo:

-   -   1) Increases the ratio of Bacteroidetes species to Firmicutes        species in fecal samples when samples at the end of study are        compared to samples at the onset of study;    -   2) Improves severity of pain and discomfort;    -   3) Increases stool consistency;    -   4) Decreases urgency to evacuate;    -   5) Decreases stool frequency;    -   6) Decreases bloating    -   7) Increases sense of complete evacuation.

Utilization of FORMULA A to treat idiopathic diarrhea such as aparasitic infection, a viral infection and a symptomatic response to afood is expected to also improve the severity of pain and discomfort,increase stool consistency, decrease the urgency to evacuate, decreasestool frequency, and decrease the sensation of bloating.

EXAMPLE 7 Case Report of FORMULA A Use in Fecal Bacteriotherapy

A 55 year old woman with chronic diarrhea that was associated with C.difficile infection initially presents to her primary care physiciandiarrhea of 8 months duration that originally started shortly followingtreatment with cephalosporin and quinolone antibiotics for back surgeryand a pulmonary infection. During these eight months, she is repeatedlytreated with metronidazole and vancomycin, and required severalhospitalizations for intravenous hydration. The patient complains ofloose small bowel movements every 15 minutes, accompanied by greaturgency and rectal tenesmus. She wears diapers at all times and loses10% of her body weight. Flexible sigmoidoscopy demonstrates classicpseudomembranous colitis. Stool samples are positive for C. difficiletoxins A and B, and stool culture confirms heavy growth of thisbacterium. The patient is again treated with vancomycin, but fails torespond. She is subsequently prescribed nitazoxanide(2-acetyloxy-N-(5-nitro-2-thiazolyl)benzamide), 500 mg orally, twicedaily. Ten day after discontinuation of Nitazoxanide, the patient hasrecurrence of her original diarrheal symptoms. Endoscopic analysisindicates return of pseudomembranous colitis, and stool studies areagain positive for C. difficile. Despite two more cycles ofNitazoxanide, including one lasting a full month, and continuousadministration of Florastor, a probiotic containing Saccharomycesboulardii, the C. difficile-induced colitis reoccurrs.

Since conventional treatment fails to resolve the C. diff, fecalbacteriotherapy is offered to break the cycle of recurrences and achievea potential cure. Informed consent is obtained from both the patient andthe donor following discussion with each of the potential risks,benefits, and alternative options. The patient is maintained onNitazoxanide until the day before the procedure. The patient's husbandof 44 years (donor) is prescribed FORMULA A 3 times a day for 2 weeks.Fecal donor material is taken from him, who has no risk factors forblood-borne communicable diseases, has no recent exposure toantibiotics, has no gastrointestinal symptoms of any kind, and testsnegative for common stool pathogens and C. difficile. Bacteriotherapy isdelivered into the patient's right colon by way of a colonoscopy. Thepatient is expected to have her first solid bowel movement on the secondday following treatment. It is expected that her abdominal paingradually subsides, and at one month following bacteriotherapy and it isexpected that her stool samples will be negative for C. difficile. Thepatient is expected to commence FORMULA A twice a day. At six monthsfollow-up, the patient is expected to report once-daily formed stools.

All measurements disclosed herein are at standard temperature andpressure, at sea level on Earth, unless indicated otherwise. Allmaterials used or intended to be used in a human being arebiocompatible, unless indicated otherwise.

The foregoing embodiments are presented by way of example only; thescope of the present invention is to be limited only by the followingclaims.

What is claimed is:
 1. A method for treating diarrhea associated with administration of metformin in a diabetic human in need thereof, comprising: a) administering to the diabetic human a pharmaceutical composition comprising a therapeutically effective amount of metformin; and b) administering to the diabetic human a therapeutic composition, comprising a therapeutically effective amount of: i. purified agave inulin; ii. β-glucan extracted from oat; and iii. blueberry pomace extract comprising at least one polyphenolic compound, to effectively treat the diarrhea in the diabetic human.
 2. The method according to claim 1, comprising at least about 3 grams purified agave inulin.
 3. The method according to claim 1, comprising at least about 2 grams β-glucan extracted from oat.
 4. The method according to claim 1, wherein the extracted β-glucan is oat (1,3/1,4) β-glucan.
 5. The method according to claim 1, wherein the blueberry pomace extract comprises at least about 700 mg total polyphenolics.
 6. The method according to claim 1, wherein the blueberry pomace extract comprises anthocyanin.
 7. The method according to claim 1, wherein the therapeutic composition comprises: i. at least about 3 grams purified agave inulin; ii. at least about 2 grams β-glucan extracted from oat; and iii. blueberry pomace extract comprising at least about 700 mg total polyphenolics.
 8. The method according to claim 1, wherein the therapeutic composition is formulated as a: medical food, beverage, suspension, powder, bar, smoothie, yogurt, shake, capsule, or tablet.
 9. The method according to claim 1, wherein said pharmaceutical composition and said therapeutic composition are formulated together as a combination and administered simultaneously.
 10. The method according to claim 1, wherein the therapeutic composition is administered orally to said diabetic human at least once per day, or at least twice per day, or at least three times per day.
 11. The method according to claim 1, wherein said diabetic human has Type 1 diabetes.
 12. The method according to claim 1, wherein said diabetic human has Type 2 diabetes.
 13. The method according to claim 1, wherein said diabetic human experiences less episodes of diarrhea when administered the therapeutic composition, as compared to the number of episodes of diarrhea experienced by said diabetic human when not administered the therapeutic composition.
 14. The method according to claim 1, wherein said diabetic human has less watery stool.
 15. A method for treating diarrhea in a diabetic human in need thereof, comprising: a) administering to the diabetic human a therapeutically effective amount of: metformin, purified agave inulin, oat glucan extract, and blueberry pomace extract, to effectively treat the diarrhea in the diabetic human.
 16. The method according to claim 15, wherein at least about 3 grams purified agave inulin is administered.
 17. The method according to claim 15, wherein the oat glucan extract is oat (1,3/1,4) β-glucan.
 18. The method according to claim 15, wherein at least about 2 grams oat glucan extract is administered.
 19. The method according to claim 15, wherein the blueberry pomace extract comprises at least one polyphenolic compound.
 20. The method according to claim 15, wherein the blueberry pomace extract comprises anthocyanin.
 21. The method according to claim 15, wherein the blueberry pomace extract comprises at least about 700 mg total polyphenolics.
 22. The method according to claim 15, wherein the purified agave inulin, oat glucan extract, and blueberry pomace extract are formulated together as a: medical food, beverage, suspension, powder, bar, smoothie, yogurt, shake, capsule, or tablet.
 23. The method according to claim 15, wherein said diabetic human has Type 1 diabetes.
 24. The method according to claim 15, wherein said diabetic human has Type 2 diabetes.
 25. The method according to claim 15, wherein said diabetic human experiences reduced episodes of diarrhea.
 26. The method according to claim 15, wherein said diabetic human experiences less watery stool.
 27. A method to reduce diarrhea in a Type 2 diabetic human taking a therapeutically effective amount of metformin, comprising: a) administering to the Type 2 diabetic human taking metformin, a therapeutically effective amount of a dietary gastrointestinal microbiome modulating composition, comprising, per dose: i. at least about 3 grams purified agave inulin; ii. at least about 2 grams oat glucan extract; and iii. blueberry pomace extract comprising at least about 700 mg total polyphenolics, wherein said Type 2 diabetic patient experiences reduced episodes of diarrhea.
 28. The method according to claim 27, wherein the oat glucan extract is oat (1,3/1,4) β-glucan.
 29. The method according to claim 27, wherein the dietary gastrointestinal microbiome modulating composition is administered orally to the Type 2 diabetic patient at least once per day, or twice per day, or three times per day.
 30. The method according to claim 27, wherein the dietary gastrointestinal microbiome modulating composition is formulated as a: medical food, beverage, suspension, powder, bar, smoothie, yogurt, shake, capsule, or tablet. 